Human mena isoforms serve as markers of epithelial to mesenchymal transition and sensitivity to egfr inhibition in human cancer cells

Abstract

The present invention discloses a method for discriminating between sensitive and resistant tumours to a treatment with EGFR inhibitor drugs comprising: in vitro testing whether tumour material expresses the hMena +11a splicing variant of hMe-na, the tumour positive to said testing being sensitive to the treatment.

1 . A method for discriminating between sensitive and resistant tumours to a treatment with EGFR inhibitor drugs comprising:
 in vitro testing whether tumour material expresses the hMena +11a  splicing variant of hMena, the tumour positive to said testing being sensitive to the treatment.   
     
     
         2 . The method according to  claim 1 , wherein the tumour is an epithelial tumour. 
     
     
         3 . The method according to  claim 1 , wherein the tumour material is selected from the group consisting of a specimen of tumour tissue, tumour cells, and in blood circulating tumour cells. 
     
     
         4 . The method according to  claim 1 , wherein the testing of the hMena +11a  expression is made by detecting the hMena +11a  isoform protein using one or more labelled antibody or labelled fragment thereof specific for said hMena +11a  isoform. 
     
     
         5 . The method according to  claim 4 , wherein said antibody or fragment thereof is a polyclonal or a monoclonal antibody specific for the peptide SEQ ID NO: 3 or SEQ ID NO: 4 or fragments thereof. 
     
     
         6 . The method according to  claim 1 , wherein the testing of the hMena +11a  expression is made by detecting the hMena +11a  mature transcription products by hybridisation with a labelled probe specific for hMena +11a , said probe having sequence selected from the sequence SEQ ID NO: 1 or a portion thereof, comprising the sequence SEQ ID NO: 2 or a portion thereof. 
     
     
         7 . The method according to  claim 1 , wherein the testing of the hMena +11a  expression is made by detecting the hMena +11a  mature transcription products by PCR amplification of the hMena +11a  variant. 
     
     
         8 . The method according to  claim 7 , wherein the amplification is made using primers amplifying a region of hMena comprising at least a portion of the sequence SEQ ID NO: 2. 
     
     
         9 . The method according to  claim 8 , wherein the amplification is made by RT-PCR using labelled primers specific for the hMena +11a  variant. 
     
     
         10 . A method for detecting in the blood circulating tumoral cells of tumours sensitive to EGFR inhibitor drugs comprising:
 in vitro testing whether the particulate portion of sample shows expression the hMena +11a  splicing variant of hMena, the portion positive to said testing comprising circulating tumoral cells;   detecting on blood samples of patients affected by tumours sensitive to treatment with EGFR inhibitor drugs the expression or non expression of the Mena +11a  variant of hMena; and   identifying the presence of said free tumoral cells when expression of the Mena +11a  variant is detected.   
     
     
         11 . A kit for identifying tumours sensitive to a treatment with EGFR inhibitor drugs or a kit for detecting in the blood free tumoral cells of tumours sensitive to EGFR inhibitor drugs comprising means for detecting the expression of hMena +11a  splicing variant, wherein said means are selected from the group consisting of: a) at least an antibody or fragment thereof specific for hMena +11a  splicing isoform, said antibody or fragments capable of being detected; b) one or more labelled probes specific for hMena +11a  mature transcription products; and c) two or more primers amplifying the hMena +11a  variant or a region of hMena +11a  comprising at least a portion of the sequence SEQ ID NO: 2. 
     
     
         12 . (canceled) 
     
     
         13 . The kit according to  claim 11 , wherein the antibody or fragment thereof is a polyclonal or monoclonal antibody specific for the sequence SEQ ID NO: 4 or its fragments. 
     
     
         14 . The kit according to  claim 11 , wherein said labelled probe has sequence SEQ ID NO: 1 or a portion thereof comprising at least part of the sequence SEQ ID NO: 2. 
     
     
         15 . The kit according to  claim 11 , wherein the primers amplify the hMena +11a  variant or a region of hMena +11a  are labelled primers. 
     
     
         16 . The kit according to  claim 1  further comprising reagents suitable for the detection of the labelled signal and optionally means for labelling nucleic acids. 
     
     
         17 . A polyclonal or monoclonal antibody specifically binding the peptides of sequence SEQ ID NO: 3 or SEQ ID NO: 4; or a probe having sequence SEQ ID NO: 5 or SEQ ID NO: 6; or a primer having sequence selected from the group consisting of SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, and SEQ ID NO: 11. 
     
     
         18 - 19 . (canceled) 
     
     
         20 . The method according to  claim 2 , wherein the epithelial tumour is selected from the group consisting of breast, colorectal, ovarian, pancreatic, and lung cancer.